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J. Biochem, 1987, Vol. 101, No. 4 1041-1049
© 1987 Japanese Biochemical Society

other

Reassembly of Nucleosomal Histone Octamers during Replication of Chromatin1

Kyo YAMASU and Tatsuo SENSHU2

Department of Biochemistry, Tokyo Metropolitan Institute of Gerontology Itabashi-ku, Tokyo 173

2 To whom correspondence should be addressed.

We have examined critically whether or not new and old histones mix in the octameric units of nucleosomes during chromatin replication. MH-134SC cells were density-labeled by culturing with amino acid mixtures enriched with dense isotopes 2H, 13C, and 15N. Mononucleosomes obtained from labeled cells were fractionated by rate zonal sedimentation through a sucrose gradient in heavy water (Senshu et al. (1985) Eur J. Biochem. 150, 575–580). The fractionation can be performed under conditions that do not destabilize nucleosomes. Density-labeling yielded heterogeneous mononucleosome species which showed higher sedimentation rates than normal mononucleosomes. However, they were indistinguishable with respect to their protein compositions, electrophoretic mobilities, electrophoretic patterns of single-stranded DNA fragments liberated by DNase I digestion, electrophoretic mobilities and sedimentation velocities of the DNA moieties, and metabolic stabilities of the histone moieties. These data suggest that the heterogeneity of density-labeled mononucleosomes resulted from the formation of histone octamers density-substituted to different degrees. This would be an inevitable consequence of mixing of new and old histones in the octameric unit of nucleosomes.

1This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.


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